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The Journal of Neuroscience, February 21, 2007, 27(8):2135-2144; doi:10.1523/JNEUROSCI.2861-06.2007

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Cellular/Molecular
Number and Density of AMPA Receptors in Individual Synapses in the Rat Cerebellum as Revealed by SDS-Digested Freeze-Fracture Replica Labeling

Miwako Masugi-Tokita,^1,2 Etsuko Tarusawa,^1,2,3 Masahiko Watanabe,⁴ Elek Molnár,⁵ Kazushi Fujimoto,⁶  and Ryuichi Shigemoto^1,2,3

¹Division of Cerebral Structure, National Institute for Physiological Sciences, Myodaiji, Okazaki 444-8787, Japan, ²Solution Oriented Research for Science and Technology, Japan Science and Technology Agency, Kawaguchi, Saitama 332-0012, Japan, ³Department of Physiological Sciences, Graduate University for Advanced Studies, Sokendai 444-8787, Japan, ⁴Department of Anatomy, Hokkaido University, Sapporo 060-8638, Japan, ⁵Medical Research Council, Centre for Synaptic Plasticity, Department of Anatomy, School of Medical Sciences, University of Bristol, Bristol BS8 1TD, United Kingdom, and ⁶Section of Physiological Anatomy, Fukui Prefectural University, Fukui 910-1195, Japan

Correspondence should be addressed to Dr. Ryuichi Shigemoto, National Institute for Physiological Sciences, Myodaiji, Okazaki 444-8787, Japan. Email: shigemot{at}nips.ac.jp

The number of AMPA receptor (AMPAR) is the major determinant of synaptic strength at glutamatergic synapses, but little is known about the absolute number and density of AMPARs in individual synapses. Using SDS-digested freeze-fracture replica labeling, which has high detection efficiency comparable with electrophysiological noise analysis for functional AMPAR, we analyzed three kinds of excitatory synapses in the molecular layer of the adult rat cerebellum. In parallel fiber (PF)–Purkinje cell (PC) synapses, we found large variability in the number (38.1 ± 34.4 particles per synapse, mean ± SD; range, 2–178 particles per synapse) and density (437 ± 277 particles/µm²; range, 48–1210 particles/µm²) of immunogold-labeled AMPARs. Two-dimensional view and high sensitivity of this method revealed irregular-shaped small AMPAR clusters within synapses. Climbing fiber (CF)–PC synapses had higher number of AMPAR labeling (68.6 ± 34.5 particles per synapse) than PF–PC and PF–interneuron synapses (36.8 ± 14.4 particles per synapse). Furthermore, AMPAR density at CF–PC and PF–interneuron synapses was approximately five times higher and more uniform than that at PF–PC synapses. These results suggest input- and target-dependent regulation of AMPAR-mediated synaptic strength.

Key words: glutamate receptor; AMPA receptor; cerebellum; Purkinje cell; synapse; SDS freeze-fracture replica labeling; 2 receptor; electron microscopy

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Received July 5, 2006; revised Jan. 17, 2007; accepted Jan. 18, 2007.

Correspondence should be addressed to Dr. Ryuichi Shigemoto, National Institute for Physiological Sciences, Myodaiji, Okazaki 444-8787, Japan. Email: shigemot{at}nips.ac.jp

This article has been cited by other articles:

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